DCL2 | Dicer-like protein 2
AS15 3100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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156.9 | 157 kDa
Nicotiana tabacum, Zea mays
50 µg of total protein from Arabidopsis thaliana whole vegetative rosette, DCL2 overexpression line (a), wild type Col-0 (b), dcl2-1 intron insertion (c), extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with laemmli buffer at 95°C/5 min. were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent and expose to Amersham Hyperfilms ECL for 120 seconds.
Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
Endoribonuclease Dicer homolog 2 is a protein which processes secondary siNAs, which is an important part in the transitive silencing of transgenes.
Alternative names: Dicer-like protein 2